The Molecular Characterization of Rhizobacteria Isolates from Saki, Nigeria

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  •   O. M. Adeoti

  •   A. T. Usman

Abstract

Plant growth promoting Rhizobacteria (PGPR) are important in the agricultural sector. Although different microorganism live in the soil but thrive in PGPR rhizosphere zones, improve the production and protection to them from diseases by production of metabolites, volatile compounds and phytohormones and Induced Systemic Resistance (ISR). This study was aimed at isolating and characterize molecularly the bacteria from the rhizosphere of pepper (Capsicum annuum), vegetable (Spinacia oleracea), rice (Oryza sativa) grown in Saki. The rhizosphere sample of pepper, rice and vegetable were collected between the hours of 1:40-2:00pm. The colonial characteristics, Gram staining techniques, biochemical test were carried out. The isolates were tested against eight antibiotics (Ceftriaxone, Gentamicin, Ceftazidime, Ofloxacin, Augmentin, Cefuroxime, Erythromycin, and Cloxacillin) by using disc diffusion method. PCR techniques and subsequent use of sanger method was used for sequencing. DNA extract was obtained using the lysozyme-SDS-phenol chloroform method in the prepared kits by Jena Bioscience. Amplification of the 16S rRNA gene was performed with the use of T1-Thermocycler PCR machine with 1492R primers pA (5′-TAC GGYBTAC CTT GTT ACG ACT T3′) and 27F primers pH (5′-AGAGTTTGATCMTGGCTCAG3'). This Primer was used for PCR amplification of 16S rRNA gene. All bacterial isolates were catalase (+) and indole (-). Antibiotics screening showed that all isolates resist most of the antibiotics except Ofloxacin and ciprofloxacin while sample 1, 2, 5 and 6was susceptible to Gentamicin. Six bacterial isolates obtained were characterized on molecular basis of 16S rRNA sequencing. The identified isolates were: Bacillus thuringensis B. weidmanii, B. cereus.


Keywords: PGPR; ISR; Rhizobacteria; Biopesticides; Biofertilizer; 16S rRNA

References

D. Sylvia, Fuhrmann J, Hartel P, Zuberer D. ‘Principles and Applications of Soil Microbiology’. Pearson Education Inc., New Jersey, 2005.

M. Ashrafuzzaman, Hossen FA, Ismail MR, Hoque MA, Islam MZ, Shahidullah SM, Meon S. ‘Efficiency of plant growth promoting rhizobacteria for the enhancement of rice growth’’. Afr J Biotechnology 8: 1247-1252, 2009.

D. Saravanakumar, Kumar CV, Kumar N, Samiyappan R ‘PGPR-induced defense responses in the tea plant against blister blight disease’’. Crop Prot 26:556–565, 2007.

Xuan Yu, Xu Liu, Tian-Hui Zhu, Guang-Hai Liu, Cui Mao. ‘Co-inoculation with phosphate-solubilizing and nitrogen-fixing bacteria on solubilization of rock phosphate and their effect on growth promotion and nutrient uptake by walnut’’. European J Soil Biol 50: 112-117, 2012.

V.O. Martinez, Jorquera M.A, Crowley DE, Gajardo G, Mora ML. ‘Mechanisms and practical considerations involved in plant growth promotion by rhizobacteria’’. J Soil Sci Plant Nutr 10: 293-319, 2010.

B.D.S. Pinheiro, E.D.M.D. Castro and C.M. Guimaraes, ‘Sustainability and profitability of aerobic rice production in Brazil’’. Field Crop Res., 97: 34–42, 2006.

S.I.A. Pereira, and P.M.L. Castro, Phosphate-solubilizing rhizobacteria enhance Zea mays growth in agricultural P-deficient soils. Ecol. Eng., 73: 526–535, 2014.

Q. Wang, F. Li, L. Zhao, E. Zhang, S. Shi, W. Zhao, W. Song and M.M. Vance, ‘Effect of irrigation and nitrogen application rates on nitrate nitrogen distribution and fertilizer nitrogen loss, wheat yield and nitrogen uptake on a recently reclaimed sandy farmland’’. Plant Soil., 337: 325–339, 2010.

M. Zaman, M.L. Nguyen and J.D. Blennerhassett, ‘The effect of different rates of urea with or without urease inhibitor (NBPT) on wheat yield and quality’’. Agric. J., 5: 309–312, 2010.

T. Emilsson, Berndtsson, J.C., Mattson, J.E. and K. Rolf, Effect of using conventional and controlled release fertilizer on nutrient runoff from various vegetated roof systems. Ecol. Eng., 29: 260– 271, 2007.

P. Chandna, M.L. Khurana, J.K. Ladha, M. Punia, R.S. Mehla and R. Gupta, Spatial and seasonal distribution of nitrate-N in groundwater beneath the rice-wheat cropping system of India: a geospatial analysis. Environ. Monit. Assess. 178: 545–562, 2011.

S.K. Rawat, R.K. Singh and R.P. Singh, ‘Seasonal variation of nitrate level in ground and surface waters of Lucknow and its remediation using certain aquatic macrophytes’’. Int. J. Lake Riv., 3: 25–35, 2010.

L. C., Van Loon and Bakker, P. A. H. M. Root-associated bacteria inducing systemic resistance. In S. S. Gnanamanickam (Ed.), Plant-associated bacteria (pp. 269–316). Dordrecht, the Netherlands: Springer, 2006.

V.V. Datar, Mayee CD Assessment of losses in tomato yield due to early blight. Indian Phytopathology 34:191–195, 1981

J.B. Jones, Jones JP, Stall RE, Zitter TA ‘Compendium of tomato diseases. APS Press’’, St. Paul, 1991.

D.O. Chellemi, Olson SM, Mitchell DJ, Seeker J, McSorley R ‘Adaptation of soil solarization to the integrated management of soil borne pests of tomato under humid conditions’’. Phytopathology 87:250–258, 1997.

N.A. Agrios (1988) Plant pathology, 3rd edn. Academic Press, U S A pp 220–222.

Y. Bashan, Kamenev, A. A., and de-Bashan, L. E. Tricalcium phosphate is inappropriate as a universal selection factor for isolating and testing phosphate-solubilizing bacteria that enhance plant growth: A proposal for an alternative procedure. Biol. Fertil. Soils 49:465-479.

R. Prasad, Kumar, M., and Varma, A. (2015). Role of PGPR in Soil Fertility and Plant Health. Plant-Growth-Promoting Rhizobacteria (PGPR) and Medicinal Plants. Berlin: Springer International Publishing, doi: 10.1007/978-3-319-13401-7_1.

P. Mehta, Walia A., Kulshrestha S., Chauhan A., Shirkot C.K. Efficiency of plant growth-promoting P solubilizing Bacillus circulans CB7 for enhancement of tomato growth Undernet house conditions. Basic Microbiol.531–1210.1002/jobm. 201300562 2014 [PubMed] [CrossRef.] [GoogleScholar.

S. Gupta, Kaushal R, Sood G, Sharma R, Kirti S. Screening of indigenous plant growth promoting rhizobacteria associated with Capsicum annuum L. in high hills temperate wet conditions of Himachal Pradesh (India). Mol Soil Biol 7 (3): 1-8, 2016.

Paula García-Fraile1, Esther Menéndez Raúl Rivas2, Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology, Academy of Sciences of the Czech Republic, Videnska 1083,14220 Praha4, Prague, Czech Republic; 2015.

E. Cachet, Barkar M., Read TD, Priest FG. 2008. B. thuringiensis strain producing a polyglutamate capsule resembling that of B. anthracis. FEMS Microbiol. Lett. 285;220 -26.

Paula García-Fraile1, Esther Menéndez Raúl Rivas2, Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology, Academy of Sciences of the Czech Republic, Videnska 1083,14220 Praha4, Prague, Czech Republic; 2015.

S. Stamenkovic, V. Beskoski, I. Karabegovic, M. Lazic, N. Nikolic Microbial fertilizers: a comprehensive review of current findings and future perspectives Span. J. Agric. Res., 16 (2018), pp. 1-18,

M. Usha Rani Arundhathi and Gopal Reddy Screening of rhizobacteria containing plant growth promoting (PGPR) traits in rhizosphere soils and their role in enhancing growth of pigeon pea (Andhra University, Visakhapatnam, India) Department of Botany. Andhra University, India 2011.

S.T. Patel, F.P. Minocheherhomji Review: plant growth promoting rhizobacteria: blessing to agriculture Int. J. Pure Appl. Biosci., 6 (2018), pp. 481-492W.

Raza, S. Yousaf, F.U. Raje Plant growth promoting activity of volatile organic compounds produced by Bio-control strains Sci. Lett., 4 (2016), pp. 40-43.

A. Beneduzi, Ambrosini, A., and Passaglia, L. M. Plant growth promoting rhizobacteria (PGPR): Their potential as antagonists and biocontrol agents. Genet. Mol. Biol. 35:1044-1051, 2012.

J.G. Holt, Krieg NG, Sneathm PHA, Staley JT and Williams ST, Bergey’s Manual of Determinative Bacteriology, 9th edn. Baltimore, MD: Williams and Williams. Potential Plant Growth-Promoting Activity of Rhizobacteria Pseudomonas spin Oryza sativa, 1994.

Gauthier 2008 The Sanger sequencing method in 6 steps 10.5424/sjar/2018161-12117.

Tamura and Nei M. (1993). Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution 10:512-526.

Kumar S., Stecher G., Li M., Knyaz C., and Tamura K. (2018). MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution 35:1547-1549.

Mahwish Z, Abbasi MK, Hameed S and Rahim N. 2015. Isolation and identification of indigenous plant growth promoting rhizobacteria from Himalayan region of Kashmir and their effect on improving growth and nutrient contents of maize (Zea mays L.). Front Microbiol 6 (207): 1-10.

R.F. Cakmakci, Erat U. G., A and M.F. Donmez. 2007. The influence of PGPR on the growth parameters, antioxidant and pentose substrate phosphate cycle enzymes in wheat and spinach plant. J. plant Nutr.Soil Sci. 170; 288-357.

K.K. Pal, K.V.B.R. Tilak, A.K. Saxena, R. Dey and C.S. Singh. Suppression of maize root diseases caused by Macrophomina phaseolina, Fusarium moniliforme and Fusarium graminearum by plant growth promoting rhizobacteria. Microbiol. Res., 156: 209-223, 2001.

Shaharoona B, Naveed M, Arshad M, Zahirm ZA. 2008. Fertilizer dependent efficiency of Pseudomonads for improving growth, yield, and nutrient use efficiency of wheat (Triticum aestivum L.). Appl Microbiol Biotechnology 79: 147-155 LettApplMicrobiol42:155–159.

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How to Cite
Adeoti, O. M., & Usman, A. T. (2021). The Molecular Characterization of Rhizobacteria Isolates from Saki, Nigeria. European Journal of Biology and Biotechnology, 2(2), 26-38. https://doi.org/10.24018/ejbio.2021.2.2.159